Brain glucose induces tolerance of Cryptococcus neoformans to amphotericin B during meningitis.

Antibiotic tolerance is the ability of a susceptible population to survive high doses of cidal drugs and has been shown to compromise therapeutic outcomes in bacterial infections. In comparison, whether fungicide tolerance can be induced by host-derived factors during fungal diseases remains largely unknown. Here, through a systematic evaluation of metabolite-drug-fungal interactions in the leading fungal meningitis pathogen, Cryptococcus neoformans, we found that brain glucose induces fungal tolerance to amphotericin B (AmB) in mouse brain tissue and patient cerebrospinal fluid via the fungal glucose repression activator Mig1. Mig1-mediated tolerance limits treatment efficacy for cryptococcal meningitis in mice via inhibiting the synthesis of ergosterol, the target of AmB, and promoting the production of inositolphosphorylceramide, which competes with AmB for ergosterol. Furthermore, AmB combined with an inhibitor of fungal-specific inositolphosphorylceramide synthase, aureobasidin A, shows better efficacy against cryptococcal meningitis in mice than do clinically recommended therapies.

Regulatory basis for reproductive flexibility in a meningitis-causing fungal pathogen.

Pathogenic fungi of the genus Cryptococcus can undergo two sexual cycles, involving either bisexual diploidization (after fusion of haploid cells of different mating type) or unisexual diploidization (by autodiploidization of a single cell). Here, we construct a gene-deletion library for 111 transcription factor genes in Cryptococcus deneoformans, and explore the roles of these regulatory networks in the two reproductive modes. We show that transcription factors crucial for bisexual syngamy induce the expression of known mating determinants as well as other conserved genes of unknown function. Deletion of one of these genes, which we term FMP1, leads to defects in bisexual reproduction in C. deneoformans, its sister species Cryptococcus neoformans, and the ascomycete Neurospora crassa. Furthermore, we show that a recently evolved regulatory cascade mediates pre-meiotic unisexual autodiploidization, supporting that this reproductive process is a recent evolutionary innovation. Our findings indicate that genetic circuits with different evolutionary ages govern hallmark events distinguishing unisexual and bisexual reproduction in Cryptococcus.

Potential mobilization of cadmium and zinc in soils spiked with smithsonite and sphalerite under different water management regimes.

Particulate cadmium (Cd) and zinc (Zn) are ubiquitous in agricultural soils of Pb-Zn mining regions. Water management serves as an important agronomic measure altering the bioavailability of Zn and Cd in soils, but how this affects particulate Cd and Zn and the underlying mechanisms remain largely unknown. Microcosm soil incubation combined with spectroscopic and microscopic characterization was conducted. During a two-year-long incubation period we observed that the concentrations of soil CaCl2-extractable Zn and Cd increased 3-10 times in sphalerite-spiked soils and 1-2 times in smithsonite-spiked soils under periodic flooding conditions due to the long-term dissolution of sphalerite (SP) and smithsonite (SM). However, the increase in the concentration of CaCl2-extractable metals (Zn: from 0.607 mg kg-1 to 1.051 mg kg-1 and Cd: from 0.047 mg kg-1 to 0.119 mg kg-1) was found only in SP-treatment under continuous flooding conditions, indicating the mobilization of metals. Ultrafiltration analysis shows that the nanoparticulate fraction of Zn and Cd in soil pore water increased 5 and 7 times in SP-treatments under continuous flooding conditions, suggesting the increment of metal pools in soil pore water. HRTEM-EDX-SAED further reveals that these nanoparticles were mainly crystalline ZnS and Zn-bearing sulfate nanoparticles in the SP-treatment and amorphous ZnCO3 and ZnS nanoparticles in the SM-treatment. Therefore, the formation of the stable crystalline Zn-bearing nanoparticles in the SP-treatment may explain the elevation of the concentration of soil CaCl2-extractable Zn and Cd under continuous flooding. The potential mobility of particulate metals should therefore be expected in scenarios of continuous flooding such as paddy soils and wetland systems.

Potential environmental risk of natural particulate cadmium and zinc in sphalerite- and smithsonite-spiked soils.

Cadmium (Cd)-bearing sphalerite and smithsonite ore particles are ubiquitous in soils near metal-mining areas. Previous studies indicate that smithsonite is more readily dissolved in acidic waters and soils than sphalerite but the mobility of Cd and zinc (Zn) derived from these ores in soils is unknown. Using microcosm incubation experiments and microscopic and spectroscopic analysis, we found that the mobility of Cd and Zn derived from smithsonite is higher than from sphalerite. The mobilization rates of Cd (16.6%) and Zn (13.7%) released from smithsonite in soils after 30-day incubation experiments were higher than those from sphalerite (Cd, ~ 1.42%; Zn, ~ 0.75%). Moreover, the percentages of Cd2+ and Zn2+ in soil pore water showed a dynamic increase in smithsonite-spiked treatments but a decrease in sphalerite-spiked treatments. HRTEM-EDX-SAED analysis further indicates the occurrence of dynamic transformation of amorphous Cd and Zn species in soil pore water to crystalline ZnS and iron oxides in sphalerite-spiked soil but crystalline ZnCO3 nanoparticles were dynamically transformed to amorphous metal-bearing species in smithsonite-spiked soil. The opposite transformation trends in pore water of Zn ore-spiked soils provide new insights into the Cd environmental risks in soils affected by Zn mining.

Prediction models for rice cadmium accumulation in Chinese paddy fields and the implications in deducing soil thresholds based on food safety standards.

Deducing soil cadmium (Cd) safety thresholds should be different for rice cultivars with different capacities to accumulate Cd to guarantee safe rice production in China. This study developed prediction models based on soil properties and deduced soil safety thresholds for Cd translocation from thirty-three paddy soils by two contrasting rice cultivars, Yelicanghua (high Cd accumulator, HCd) and Longhuamaohu (low Cd accumulator, LCd). A total of 330 paired field validation samples were used to examine the accuracy of prediction models and soil safety thresholds. The average soil Cd concentration was 0.26 (range 0.057-0.72) mg kg-1. The average brown rice Cd concentrations were 0.14 (0.043-0.55) mg kg-1 in HCd and 0.024 (0.007-0.15) mg kg-1 in LCd in 2017, with corresponding values of 0.16 (0.016-0.66) and 0.027 (0.009-0.10) mg kg-1 in 2018. Soil total Cd and pH were the two most important variables exhibiting direct effects on Cd concentrations in HCd, explaining 66% of the variance across the 33 soils. Soil total Cd, pH and organic carbon (OC) were the three most important variables in LCd, explaining 75% of the variance. Soil safety thresholds ranged from 0.27 to 1.00 mg kg-1 for HCd and from 4.52 to 46.9 mg kg-1 for LCd with pH ranging from 4.5 to 8.0. The validation results suggest ∼60% for HCd or the current soil quality standard (SQS) and 88% for LCd of the validation samples were suitable to meet the food quality standard (FQS), with 6.4% and 12%, respectively, of the validation soils unsuitable for rice cultivation. The current Chinese SQS is too strict for LCd which may be grown safely in moderately polluted soils and the derivation of soil thresholds should therefore consider the abilities of different cultivars to accumulate Cd.

Effects of a natural sepiolite bearing material and lime on the immobilization and persistence of cadmium in a contaminated acid agricultural soil.

Soil contamination with cadmium (Cd) represents a substantial threat to human health and environmental quality. Long-term effectiveness and persistence of remediation are two important criteria for the evaluation of amendment techniques used to remediate soils polluted with potentially toxic metals. In the current study, we investigated the remediation persistence of a natural sepiolite bearing material (NSBM, containing 15% sepiolite) and ground limestone (equivalent to > 98.0% CaO) on soil pH, Cd bioavailability, and Cd accumulation by pak choi (Brassica chinensis L.) during the growth of four consecutive crops in a Cd-contaminated acid soil with different amounts of NSBM (0, 0.2, 0.5, 1, 2, and 5%). Soil pH levels ranged from 5.21 to 7.76 during the first crop, 4.30 to 7.34 during the second, 4.23 to 7.80 during the third, and 4.33 to 6.98 during the fourth, and increased significantly with increasing the application rate of NSBM. Soil CaCl2-Cd and shoot Cd concentrations decreased by 8.11 to 99.2% and 6.58 to 94.5%, respectively, compared with the control throughout the four cropping seasons. A significant negative correlation was found between soil CaCl2-Cd and soil pH. Combined use of 0.1% lime and NSBM showed greater effects than NSBM alone, especially, when the application rate of NSBM was ˂ 2%. Moreover, pak choi tissue Cd concentrations in the treatments with NSBM addition alone at ≥ 2% or at ≥ 1% NSBM combined with 0.1% lime met the maximum permissible concentration (MPC) over the four crops, allowed by the Chinese and European regulations. Based on the present study, safe crop production in the test soil is possible at a soil pH > 6.38 and CaCl2-Cd < 14 µg kg-1, and soil Cd immobilization by NSBM without or with lime is a potentially feasible method of controlling the transfer of soil Cd into the food chain.

Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide.

Bacterial quorum sensing is a well-characterized communication system that governs a large variety of collective behaviours. By comparison, quorum sensing regulation in eukaryotic microbes remains poorly understood, especially its functional role in eukaryote-specific behaviours, such as sexual reproduction. Cryptococcus neoformans is a prevalent fungal pathogen that has two defined sexual cycles (bisexual and unisexual) and is a model organism for studying sexual reproduction in fungi. Here, we show that the quorum sensing peptide Qsp1 serves as an important signalling molecule for both forms of sexual reproduction. Qsp1 orchestrates various differentiation and molecular processes, including meiosis, the hallmark of sexual reproduction. It activates bisexual mating, at least in part through the control of pheromone, a signal necessary for bisexual activation. Notably, Qsp1 also plays a major role in the intercellular regulation of unisexual initiation and coordination, in which pheromone is not strictly required. Through a multi-layered genetic screening approach, we identified the atypical zinc finger regulator Cqs2 as an important component of the Qsp1 signalling cascade during both bisexual and unisexual reproduction. The absence of Cqs2 eliminates the Qsp1-stimulated mating response. Together, these findings extend the range of behaviours governed by quorum sensing to sexual development and meiosis.

Influence of nitrogen form on the phytoextraction of cadmium by a newly discovered hyperaccumulator Carpobrotus rossii.

Using hyperaccumulator plants is an important method to remove heavy metals from contaminated land. Carpobrotus rossii, a newly found Cd hyperaccumulator, has shown potential to remediate Cd-contaminated soils. This study examined the effect of nitrogen forms on Cd phytoextraction by C. rossii. The plants were grown for 78 days in an acid soil spiked with 20 mg Cd kg(-1) and supplied with (NH4)2SO4, Ca(NO3)2, urea, and chicken manure as nitrogen (N) fertilizers. Nitrification inhibitor dicyandiamide (DCD) was applied to maintain the ammonium (NH4(+)) form. Nitrogen fertilization increased shoot biomass but decreased root biomass with the highest shoot biomass occurring in the manure treatment. Compared to the no-N control, urea application did not affect shoot Cd concentration, but increased Cd content by 17% due to shoot biomass increase. Chicken manure significantly decreased CaCl2-extractable Cd in soil, and the Cd concentration and total Cd uptake in the plant. Rhizosphere pH was the highest in the manure treatment and the lowest in the NH4(+) treatments. The manure and nitrate (NO3(-)) treatments tended to have higher rhizosphere pH than their respective bulk soil pH, whereas the opposite was observed for urea and NH4(+) treatments. Furthermore, the concentrations of extractable Cd in soil and Cd in the plant correlated negatively with rhizosphere pH. The study concludes that urea significantly enhanced the Cd phytoaccumulation by C. rossii while chicken manure decreased Cd availability in soil and thus the phytoextraction efficiency.

Long-term field phytoextraction of zinc/cadmium contaminated soil by Sedum plumbizincicola under different agronomic strategies.

In two long-term field experiments the zinc (Zn)/cadmium (Cd) hyperaccumulator Sedum plumbizincicola (S. plumbizincicola) was examined to optimize the phytoextraction of metal contaminated soil by two agronomic strategies of intercropping with maize (Zea mays) and plant densities. Soil total Zn and Cd concentrations decreased markedly after long-term phytoextraction. But shoot biomass and Cd and Zn concentrations showed no significant difference with increasing remediation time. In the intercropping experiment the phytoremediation efficiency in the treatment "S. plumbizincicola intercropped with maize" was higher than in S. plumbizincicola monocropping, and Cd concentrations of corn were below the maximum national limit. In the plant density experiment the phytoremediation efficiency increased with increasing plant density and 440,000 plants ha(-1) gave the maximum rate. These results indicated that S. plumbizincicola at an appropriate planting density and intercropped with maize can achieve high remediation efficiency to contaminated soil without affecting the cereal crop productivity. This cropping system combines adequate agricultural production with soil heavy metal phytoextraction.

Effects of water management on arsenic and cadmium speciation and accumulation in an upland rice cultivar.

Pot and field experiments were conducted to investigate the effects of water regimes on the speciation and accumulation of arsenic (As) and cadmium (Cd) in Brazilian upland rice growing in soils polluted with both As and Cd. In the pot experiment constant and intermittent flooding treatments gave 3-16 times higher As concentrations in soil solution than did aerobic conditions but Cd showed the opposite trend. Compared to arsenate, there were more marked changes in the arsenite concentrations in the soil solution as water management shifted, and therefore arsenite concentrations dominated the As speciation and bioavailability in the soil. In the field experiment As concentrations in the rice grains increased from 0.14 to 0.21 mg/kg while Cd concentrations decreased from 0.21 to 0.02 mg/kg with increasing irrigation ranging from aerobic to constantly flooding conditions. Among the various water regimes the conventional irrigation treatment produced the highest rice grain yield of 6.29 tons/ha. The As speciation analysis reveals that the accumulation of dimethylarsinic acid (from 11.3% to 61.7%) made a greater contribution to the increase in total As in brown rice in the intermittent and constant flooding treatments compared to the intermittent-aerobic treatment. Thus, water management exerted opposite effects on Cd and As speciation and bioavailability in the soil and consequently on their accumulation in the upland rice. Special care is required when irrigation regime methods are employed to mitigate the accumulation of metal(loid)s in the grain of rice grown in soils polluted with both As and Cd.

Phytoextraction of Cadmium and Zinc By Sedum plumbizincicola Using Different Nitrogen Fertilizers, a Nitrification Inhibitor and a Urease Inhibitor.

Cadmium (Cd) and zinc (Zn) phytoavailability and their phytoextraction by Sedum plumbizincicola using different nitrogen fertilizers, nitrification inhibitor (dicyandiamide, DCD) and urease inhibitor (N-(n-Butyl) thiophosphoric triamide, NBPT) were investigated in pot experiments where the soil was contaminated with 0.99 mg kg(-1) of Cd and 241 mg kg(-1) Zn. The soil solution pH varied between 7.30 and 8.25 during plant growth which was little affected by the type of N fertilizer. The (NH4)2SO4+DCD treatment produced higher NH4+-N concentrations in soil solution than the (NH4)2SO4 and NaNO3 treatment which indicated that DCD addition inhibited the nitrification process. Shoot Cd and Zn concentrations across all treatments showed ranges of 52.9-88.3 and 2691-4276 mg kg(-1), respectively. The (NH4)2SO4+DCD treatment produced slightly higher but not significant Cd and Zn concentrations in the xylem sap than the NaNO3 treatment. Plant shoots grown with NaNO3 had higher Cd concentrations than (NH4)2SO4+DCD treatment at 24.0 and 15.4 mg kg(-1), respectively. N fertilizer application had no significant effect on shoot dry biomass. Total Cd uptake in the urea+DCD treatment was higher than in the control, urea+NBPT, urea+NBPT+DCD, or urea treatments, by about 17.5, 23.3, 10.7, and 25.1%, respectively.

Remediation of cadmium- and lead-contaminated agricultural soil by composite washing with chlorides and citric acid.

Composite washing of cadmium (Cd)- and lead (Pb)-contaminated agricultural soil from Hunan province in China using mixtures of chlorides (FeCl3, CaCl2) and citric acid (CA) was investigated. The concentrations of composite washing agents for metal removal were optimized. Sequential extraction was conducted to study the changes in metal fractions after soil washing. The removal of two metals at optimum concentration was reached. Using FeCl3 mixed with CA, 44% of Cd and 23% of Pb were removed, and 49 and 32% by CaCl2 mixed with CA, respectively. The mechanism of composite washing was postulated. A mixture of chlorides and CA enhanced metal extraction from soil through the formation of metal-chloride and metal-citrate complexes. CA in extract solutions promoted the formation of metal-chloride complexes and reduced the solution pH. Composite washing reduced Cd and Pb in Fe-Mn oxide forms significantly. Chlorides and CA exerted a synergistic effect on metal extraction during composite washing.

Repeated phytoextraction of four metal-contaminated soils using the cadmium/zinc hyperaccumulator Sedum plumbizincicola.

A cadmium/zinc hyperaccumulator extracted metals from four contaminated soils over three years in a glasshouse experiment. Changes in plant metal uptake and soil total (aqua regia-extractable) and available metals were investigated. Plant Cd concentrations in a high-Cd acid soil and plant Zn concentrations in two acid soils decreased during repeated phytoextraction and were predicted by soil available metal concentrations. However, on repeated phytoextraction, plant Cd concentrations remained constant in lightly Cd-polluted acid soils, as did plant Cd and Zn in alkaline soils, although soil available metal concentrations decreased markedly. After phytoextraction acid soils showed much higher total metal removal efficiencies, indicating possible suitability of phytoextraction for acid soils. However, DGT-testing, which takes soil metal re-supply into consideration, showed substantial removal of available metal and distinct decreases in metal supply capacity in alkaline soils after phytoextraction, suggesting that a strategy based on lowering the bioavailable contaminant might be feasible.

Copper changes the yield and cadmium/zinc accumulation and cellular distribution in the cadmium/zinc hyperaccumulator Sedum plumbizincicola.

Non-accumulated metals in mixed metal contaminated soils may affect hyperaccumulator growth and metal accumulation and thus remediation efficiency. Two hydroponics experiments were conducted to investigate the effects of copper (Cu) on cadmium (Cd) and zinc (Zn) accumulation by the Cd/Zn hyperaccumulator Sedum plumbizincicola, Cu toxicity and plant detoxification using chemical sequential extraction of metals, sub-cellular separation, micro synchrotron radiation based X-ray fluorescence, and transmission electron microscopy. Compared with the control (0.31 µM Cu), 5-50 µM Cu had no significant effect on Cd/Zn accumulation, but Cu at 200 µM induced root cell plasmolysis and disordered chloroplast structure. The plants held Cu in the roots and cell walls and complexed Cu in insoluble forms as their main detoxification mechanisms. Exposure to 200 µM Cu for 4 days inhibited plant Cd uptake and translocation but did not affect Zn concentrations in roots and stems. Moreover, unloading of Cd and Zn from stem to leaf was restrained compared to control plants, perhaps due to Cu accumulation in leaf veins. Copper may thus interfere with root Cd uptake and restrain Cd/Zn unloading to the leaves. Further investigation of how Cu affects plant metal uptake may help elucidate the Cd/Zn hyper-accumulating mechanisms of S. plumbizincicola.

Water management affects arsenic and cadmium accumulation in different rice cultivars.

Paddy rice (Oryza sativa L.) is a staple food and one of the major sources of dietary arsenic (As) and cadmium (Cd) in Asia. A field experiment was conducted to investigate the effects of four water management regimes (aerobic, intermittent irrigation, conventional irrigation and flooding) on As and Cd accumulation in seven major rice cultivars grown in Zhejiang province, east China. With increasing irrigation from aerobic to flooded conditions, the soil HCl-extractable As concentrations increased significantly and the HCl-extractable Cd concentrations decreased significantly. These trends were consistent with the As and Cd concentrations in the straw, husk and brown rice. Water management both before and after the full tillering stage affected As and Cd accumulation in the grains. The intermittent and conventional treatments produced higher grain yields than the aerobic and flooded treatments. Cd concentrations in brown rice varied 13.1-40.8 times and As varied 1.75-8.80 times among the four water management regimes. Cd and As accumulation in brown rice varied among the rice cultivars, with Guodao 6 (GD6) was a low Cd but high-As-accumulating cultivar while Indonesia (IR) and Yongyou 9 (YY9) were low As but high-Cd-accumulating cultivars. Brown rice Cd and As concentrations in the 7 cultivars were significantly negatively correlated. The results indicate that As and Cd accumulated in rice grains with opposite trends that were influenced by both water management and rice cultivar. Production of 'safe' rice with respect to As and Cd might be possible by balancing water management and rice cultivar according to the severity of soil pollution.

Nitrate facilitates cadmium uptake, transport and accumulation in the hyperaccumulator Sedum plumbizincicola.

The aims of this study are to investigate whether and how the nitrogen form (nitrate (NO3 (-)) versus ammonium (NH4 (+))) influences cadmium (Cd) uptake and translocation and subsequent Cd phytoextraction by the hyperaccumulator species Sedum plumbizincicola. Plants were grown hydroponically with N supplied as either NO3 (-) or NH4 (+). Short-term (36 h) Cd uptake and translocation were determined innovatively and quantitatively using a positron-emitting (107)Cd tracer and positron-emitting tracer imaging system. The results show that the rates of Cd uptake by roots and transport to the shoots in the NO3 (-) treatment were more rapid than in the NH4 (+) treatment. After uptake for 36 h, 5.6 (0.056 µM) and 29.0 % (0.290 µM) of total Cd in the solution was non-absorbable in the NO3 (-) and NH4 (+) treatments, respectively. The local velocity of Cd transport was approximately 1.5-fold higher in roots (3.30 cm h(-1)) and 3.7-fold higher in shoots (10.10 cm h(-1)) of NO3 (-)- than NH4 (+)-fed plants. Autoradiographic analysis of (109)Cd reveals that NO3 (-) nutrition enhanced Cd transportation from the main stem to branches and young leaves. Moreover, NO3 (-) treatment increased Cd, Ca and K concentrations but inhibited Fe and P in the xylem sap. In a 21-day hydroponic culture, shoot biomass and Cd concentration were 1.51 and 2.63 times higher in NO3 (-)- than in NH4 (+)-fed plants. We conclude that compared with NH4 (+), NO3 (-) promoted the major steps in the transport route followed by Cd from solution to shoots in S. plumbizincicola, namely its uptake by roots, xylem loading, root-to-shoot translocation in the xylem and uploading to the leaves. S. plumbizincicola prefers NO3 (-) nutrition to NH4 (+) for Cd phytoextraction.

[Genetic transformation of the fungus Gliocladium sp. mediated by Agrobacterium tumefaciens].

OBJECTIVE: To construct a transformation system in Gliocladium sp. 6.102, a Cordyceps-colonizing fungus producing a variety of epipolythiodioxopiperazine (ETP) compounds with drug potentials. METHODS: Agrobacterium tumefaciens-mediated transformation (ATMT) was used to transform Gliocladium sp. 6.102. The factors of bacterial cell concentration, co-cultivation time, pH and acetosyringone concentration were optimized. RESULTS; A total of 50 -100 transformants per 10(6) fungal conidia was obtained via the optimal ATMT method. The genes encoding hygromycin B phosphotase and enhanced green fluorescent protein (EGFP) were transferred into Gliocladium sp. by the optimal ATMT method. The marker genes were successfully expressed and stably maintained in the transgenic fungus. CONCLUSION: A transformation system was established for Gliocladium sp. 6.102 and this system may be useful to identify ETP biosynthetic genes in Gliocladium.

Interaction of cadmium and zinc on accumulation and sub-cellular distribution in leaves of hyperaccumulator Potentilla griffithii.

Potentilla griffithii Hook is a newly found hyperaccumulator plant capable of high tolerance and accumulation of Zn and Cd. We investigated the interactive effects between Cd and Zn on accumulation and vacuolar sequestration in P. griffithii. Stimulatory effect of growth was noted at 0.2 mM Cd and 1.25 and 2.5 mM Zn tested. Accumulation of Zn and Cd in roots, petioles and leaves were increased significantly with addition of these metals individually. However, the Zn supplement decreased root Cd accumulation but increased the concentration of Cd in petioles and leaves. The results from sub-cellular distribution showed that up to 94% and 70% of the total Zn and Cd in the leaves were present in the protoplasts, and more than 90% Cd and Zn in the protoplasts were localized in the vacuoles. Nearly, 88% and 85% of total Cd and Zn were extracted in the cell sap of the leaves suggesting that most of the Cd and Zn in the leaves were available in soluble form. The present results indicate that Zn supplement significantly enhanced the petiole accumulation of Cd and further vacuolar sequestration plays an important role in tolerance, detoxification and hyperaccumulation of these metals in P. griffithii.

Cadmium tolerance of carbon assimilation enzymes and chloroplast in Zn/Cd hyperaccumulator Picris divaricata.

To better understand the photosynthesis under stress, the effect of cadmium on carbon assimilation and chloroplast ultrastructure of a newly found Zn/Cd hyperaccumulator Picris divaricata in China was investigated in solution culture. The shoot and root Cd concentrations increased with increase in Cd supply, reaching maxima of 1109 and 5604mgkg(-1) dry weight at 75microM Cd, respectively. As Cd supply to P. divaricata increased, the shoot and root dry weight, leaf water content (except 75microM Cd), concentrations of chlorophyll a and b, chlorophyll a/b ratio and the concentration of carotenoids were not depressed at high Cd. However, the stomatal conductance, transpiration rate, net photosynthetic rate and intercellular CO(2) concentration were significantly affected when the Cd concentration reached 10, 10, 25 and 75microM, respectively. Meanwhile, carbonic anhydrase (CA; EC 4.2.1.1) activity and Rubisco (EC 4.1.1.39) content reached maxima in the presence of 50 and 5microM Cd, respectively. In addition, CA activity correlated positively with shoot Cd in plants treated with Cd at a range of 0-50microM. Moreover, the activities of NADP(+)-glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13), Rubisco and fructose-1, 6-bisphosphatase (EC 3.1.3.11) were not significantly suppressed by increased Cd supply. Although the mesophyll cell size was reduced, chloroplast ultrastructure remained intact at the highest Cd treatment. Our finding revealed that P. divaricata chloroplast and the enzymes of carbon assimilation tolerate high levels of Cd, demonstrating its potential in possible application in phytoremediation.

Antioxidative response to Cd in a newly discovered cadmium hyperaccumulator, Arabis paniculata F.

A hydroponic experiment was carried out to study the effect of cadmium (Cd) on growth, Cd accumulation, lipid peroxidation, reactive oxygen species (ROS) content and antioxidative enzymes in leaves and roots of Arabis paniculata F., a new Cd hyperaccumuator found in China. The results showed that 22-89 microM Cd in solution enhanced the growth of A. paniculata after three weeks, with 21-27% biomass increase compared to the control. Cd concentrations in shoots and roots increased with increasing Cd supply levels, and reached a maximum of 1662 and 8670 mg kg(-1) Cd dry weight at 178 microM Cd treatment, respectively. In roots, 22-89 microM Cd reduced the content of malondialdehyde (MDA), superoxide (O(2)(-1)) and H(2)O(2) as well as the activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX) and glutathione reductase (GR). In leaves, the contents of MDA, O(2)(-1) and H(2)O(2) remained unaffected by 22-89 microM Cd, while 178 microM Cd treatment significantly increased the MDA content, 69.5% higher than that of the control; generally, the activities of SOD, catalase (CAT), GPX and APX showed an increasing pattern with increasing Cd supply levels. Our present work concluded that A. paniculata has a great capability of Cd tolerance and accumulation. Moderate Cd treatment (22-89 microM Cd) alleviated the oxidative stress in roots, while higher level of Cd addition (178 microM) could cause an increasing generation of ROS, which was effectively scavenged by the antioxidative system.